LOCUS       sequence-342874-        4454 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible gene expression, pET origin, VanRAM repressor, PVanCC 
            promoter, mRFP, Carbenicillin Resistance Marker. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4454)
  TITLE     Reisch Lab Duet Plasmids
REFERENCE   2  (bases 1 to 4454)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pEVR4
FEATURES             Location/Qualifiers
     source          1..4454
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(28..46)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      42..89
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      126..581
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(213..232)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     423..444
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(696..1487,1488..1556))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIHFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1319..1338
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1557..1649)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1730..2318
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2219..2238
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2636..2658
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2650..2667
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2651..2667
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_RNA        3637..3686
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             3731..3742
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3749..4426
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
ORIGIN
        1 gctttccagt cgggaaacct gctgccaccg ctgagcaata actagcataa ccccttgggg
       61 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga
      121 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
      181 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
      241 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
      301 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
      361 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
      421 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
      481 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
      541 atttaacgcg aattttaaca aaatattaac gtttacaatt tctggcggca cgatggcatg
      601 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
      661 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
      721 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
      781 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
      841 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
      901 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
      961 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     1021 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     1081 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     1141 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     1201 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     1261 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     1321 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     1381 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     1441 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     1501 aggcaaaatg ccgcaaaaaa gtgaataagg gcgacacgga aatgttgaat actcatactc
     1561 ttcctttttc aatcatgatt gaagcattta tcagggttat tgtctcatga gcggatacat
     1621 atttgaatgt atttagaaaa ataaacaaat aggtcatgac caaaatccct taacgtgagt
     1681 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     1741 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     1801 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     1861 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     1921 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     1981 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2041 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2101 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2161 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2221 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2281 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2341 tacggttcct ggccttttgc tggccttttc gtttaagggc accaataact gccttaaaaa
     2401 aattacgccc cgccctgcca ctcatcgcag tacggcctat tggttaaaaa atgagctgat
     2461 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt ccattcgcca
     2521 ttcaggctgc gcaactgttt ggaagggcga tcggtgcggg cctcttcgct attacgccag
     2581 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     2641 tcacgacgtt gtaaaacgac ggccagtgaa acagacgaag aatccatggt caatctgcac
     2701 gaattgacca tgctgcaccc agcggtgcgc ctgcgcttgc tgctgcttca aaaacttttg
     2761 cattacgaat tgctgccagt gcatgatcac gcataatacg ttctgcacct tcggcatcac
     2821 cacagctaac tgcatccaga actgcctgat gctgacgatg tgctgccagc agatgttcat
     2881 attcggcaga caggtccatc agatccaggg ccagtgcacc ggctgctgca aacggttcaa
     2941 aaccattacg tgccagtgcg ctttcaactg caccattacc tgctgcgcta accagggtat
     3001 catgaaatgc ctgattatat gcggcataac gatccagatc ttcaccattc aggcgaccgg
     3061 ctgcaaacag tgcttcacct tctgcaatca gtacaacaaa acgtgcatgg gtttctgagg
     3121 tcataccacg ttctgccaga cgacgtgctg caaaaccttc cagaacacca cgaacttcaa
     3181 ttgcatcacg aatctgatcg ctgctaacac cacgggctgc ataaccacgt gcacccagac
     3241 gaacaaccag accttcttgt tccagtgaac gcagtgcgat acgaaccggc atacggctaa
     3301 cacccagtgc tgctgcggtc ggaatttctg caatacgttc accacttttg atttcaccgc
     3361 ttgcaatcat tttacgcagt gccatcataa cacgctgacc cggtttaata cgaggcatgt
     3421 ccatgtctat cctccttatt tacgttcgtt agtttaagcg ggcgctatca tgccataccg
     3481 cgaaaggttt tgcgccattc gatggcgcgc cgccaattat tgaaggcctc cctaacgggg
     3541 ggcctttttt tgtttctggt ctcccgctta acgatcgttg gctgattgga tccaattgac
     3601 agctagctca gtcctaggta ccattggatc caatagctgt caccggatgt gctttccggt
     3661 ctgatgagtc cgtgaggacg aaacagcctc tacaaataat tttgtttaag aactgggaag
     3721 agcgcagatt aaagaggaga aatactagat ggcttcctcc gaagacgtta tcaaagagtt
     3781 catgcgtttc aaagttcgta tggaaggttc cgttaacggt cacgagttcg aaatcgaagg
     3841 tgaaggtgaa ggtcgtccgt acgaaggtac ccagaccgct aaactgaaag ttaccaaagg
     3901 tggtccgctg ccgttcgctt gggacatcct gtccccgcag ttccagtacg gttccaaagc
     3961 ttacgttaaa cacccggctg acatcccgga ctacctgaaa ctgtccttcc cggaaggttt
     4021 caaatgggaa cgtgttatga acttcgaaga cggtggtgtt gttaccgtta cccaggactc
     4081 ctccctgcaa gacggtgagt tcatctacaa agttaaactg cgtggtacca acttcccgtc
     4141 cgacggtccg gttatgcaga aaaaaaccat gggttgggaa gcttccaccg aacgtatgta
     4201 cccggaagac ggtgctctga aaggtgaaat caaaatgcgt ctgaaactga aagacggtgg
     4261 tcactacgac gctgaagtta aaaccaccta catggctaaa aaaccggttc agctgccggg
     4321 tgcttacaaa accgacatca aactggacat cacctcccac aacgaagact acaccatcgt
     4381 tgaacagtac gaacgtgctg aaggtcgtca ctccaccggt gcttaataac gctgatagtg
     4441 ctagtgtaga tcgc
//
